Ortho-proteogenomics: multiple proteomes investigation through orthology and a new MS-based protocol.
The progress in sequencing technologies irrigates biology with an ever-increasing number of genome sequences. In most cases, the gene repertoire is predicted in silico and conceptually translated into proteins. As recently highlighted, the predicted genes exhibit frequent errors, particularly in start codons, with a serious impact on subsequent biological studies. A new "ortho-proteogenomic" approach is presented here for the annotation refinement of multiple genomes at once. It combines comparative genomics with an original proteomic protocol that allows the characterization of both N-terminal and internal peptides in a single experiment. This strategy was applied to the Mycobacterium genus with Mycobacterium smegmatis as the reference, and identified 946 distinct proteins, including 443 characterized N termini. These experimental data allowed the correction of 19% of the characterized start codons, the identification of 29 proteins missed during the annotation process, and the curation, thanks to comparative genomics, of 4328 sequences of 16 other Mycobacterium proteomes.
Gallien
Sébastien
S
Laboratoire de Spectrométrie de Masse Bio-Organique, IPHC-DSA, ULP, CNRS, UMR7178, 67 087 Strasbourg, France. sgallien@chimie.u-strasbg.fr
Perrodou
Emmanuel
E
Carapito
Christine
C
Deshayes
Caroline
C
Reyrat
Jean-Marc
JM
Van Dorsselaer
Alain
A
Poch
Olivier
O
Schaeffer
Christine
C
Lecompte
Odile
O
D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification; D018387:Codon, Initiator / Q000235:genetics; D016680:Genome, Bacterial; D023281:Genomics / Q000379:methods*; D013058:Mass Spectrometry; D009161:Mycobacterium / Q000737:chemistry / Q000235:genetics; D020102:Mycobacterium smegmatis / Q000737:chemistry / Q000235:genetics; D010446:Peptide Fragments / Q000235:genetics / Q000302:isolation & purification; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D016415:Sequence Alignment; D013045:Species Specificity
D001426:Bacterial Proteins; D018387:Codon, Initiator; D010446:Peptide Fragments; D020543:Proteome; D012329:RNA, Bacterial